A new, simple, safe, accurate and reproducible spectrophotometric analytical method was developed for the quantitative estimation of gatifloxacin in solid dosage form by mixed hydrotropic agents. Various organic solvents like methanol, chloroform, ethanol, acetonitrile, hexane, toluene are widely used to conduct the spectrophotometric analysis, But higher cost and toxicity prevents their frequent use. The enhancement of solubility of gatifloxacin was more than 4 fold in one blend containing Propylene glycol, PEG400, PEG4000 and Sodium citrate (10% w/v each) as compared to solubility in distilled water. In the present study mixed blend were employed for the spectrophotometric estimation of gatifloxacin at 288nm. The results of the analysis were validated statistically and by recovery studies. The drug follows Beer’s law in concentration range of 4-20 mcg/ml. The percent label claims and percent recoveries estimated were close to 100 with low values of standard deviation. Thus the statistical data proved the accuracy, reproducibility and precision of the proposed method. The mixed solvency concept used in the present study did not interfere in the analysis.